LOCUS       sequence-334500-        6018 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A single-component optogenetic tool for photoswitchable pyroptosis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6018)
  AUTHORS   He L, Huang Z, Huang K, Chen R, Nguyen NT, Wang R, Cai X, Huang Z, 
            Siwko S, Walker JR, Han G, Zhou Y, Jing J
  TITLE     Optogenetic Control of Non-Apoptotic Cell Death.
  JOURNAL   Adv Sci (Weinh). 2021 May 6;8(13):2100424. doi: 
            10.1002/advs.202100424. eCollection 2021 Jul.
  PUBMED    34540558
REFERENCE   2  (bases 1 to 6018)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Adv Sci 
            (Weinh)."; date: "2021-05-6"; volume: "8(13):2100424. doi"; pages: "
            10.1002/advs.202100424. eCollection 2021 Jul"
COMMENT     Sequence Label: LiPOP2
FEATURES             Location/Qualifiers
     source          1..6018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1908..1917
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1914..2624
                     /codon_start=1
                     /product="derivative of the monomeric red fluorescent 
                     protein mCherry"
                     /label=mCherry2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFNWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQCRTMGWEASTERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVDIKLDILSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2059..2077)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    2746..2867
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2783..2802)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2837..2856
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2874..3329)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3011..3032)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3223..3242
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3356..3460
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3462..3819
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3462..3653
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3485..3505)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3670..3805
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3854..4648
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3854..4648
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3908..3927)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4518..4537
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4836..4855)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4880..4927
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5256..5844
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5745..5764
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatgc
      541 catcggcctt tgagaaagtg gtcaagaatg tgatcaagga ggtaagcggc agcagaggcg
      601 atctcattcc ggtggacagc ctgcggaact ccaccagctt caggccctac tgccttctga
      661 acaggaaatt ttcaagctca aggttctgga aaccccgtta ttcatgtgtc aacctgtcaa
      721 tcaaggacat cctggagccc agtgctccag aaccagaacc ggagtgtttt ggctccttca
      781 aagtctctga tgtcgtcgat gggaacattc agggcagagt gatgttgtca ggcatgggag
      841 aagggaaaat ttctggtggg gctgcagtgt ctgacagttc cagtgcctcc atgaatgtgt
      901 gtatactgcg tgtgactcag aagacctggg agaccatgca gcatgaagcg caccttcagc
      961 agcctgagaa cgcaatcctg caacagcttg cgagtgctgg ggatgacctg tttgtggtga
     1021 ccgaggtgct gcagacaaag gaggaagtgc agatcactga ggtccacagc caagagggct
     1081 caggccagtt tacgctgcct ggagctttat gcttgaaggg tgaaggcaag ggccaccaaa
     1141 gccggaagaa gatggtgacc attcctgcag gcagcatcct ggcattccga gtggcccaac
     1201 tgcttattgg ctctaaatgg gatatccttc tcgtctcaga tgagaaacag aggacctttg
     1261 agccctcctc aggtgacaga aaagcagtgg gccagaggca ccatggcctc aatgtgcttg
     1321 ctgcgctttg ttccatcgga aagcagctca gtctcctgtc agatcaagct tccgggtctg
     1381 gtgggggtct agcttccacc atgactgagc atgtccgaga tgctgccgag agagagggag
     1441 tcatgctgat taagaaaact gcagaaaata ttgatgaggc ggcaaaagaa cttggtgggg
     1501 gatccggtgg atccggaggt ggtttggcta ctacacttga acgtattgag aagaactttg
     1561 tcattactga cccaagattg ccagataatc ccattatatt cgcgtccgat agtttcttgc
     1621 agttgacaga atatagccgt gaagaaattt tgggaagaaa ctgcaggttt ctacaaggtc
     1681 ctgaaactga tcgcgcgaca gtgagaaaaa ttagagatgc catagataac caaacagagg
     1741 tcactgttca gctgattaat tatacaaaga gtggtaaaaa gttctggaac ctctttcact
     1801 tgcagcctat gcgagatcag aagggagatg tccagtactt tattggggtt cagttggatg
     1861 gacgaattct gcagtcgacg gtaccgcggg cccgggatcc accggtcgcc accatggtga
     1921 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     1981 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     2041 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     2101 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     2161 acatccccga ctacttgaag ctgtccttcc ccgagggctt caattgggag cgcgtgatga
     2221 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     2281 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcagt
     2341 gtcgtaccat gggctgggag gcctccactg agcggatgta ccccgaggac ggcgccctga
     2401 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     2461 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcgacatca
     2521 agttggacat cctttcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     2581 agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaagcggcc gcgactctag
     2641 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     2701 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     2761 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2821 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     2881 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2941 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     3001 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3061 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     3121 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     3181 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     3241 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     3301 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     3361 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3421 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     3481 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3541 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3601 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3661 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3721 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3781 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     3841 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3901 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3961 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4021 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4081 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4141 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4201 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4261 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4321 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4381 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4441 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4501 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4561 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4621 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4681 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4741 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4801 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     4861 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     4921 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     4981 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5041 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     5101 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5161 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5221 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5281 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5341 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5401 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5461 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5521 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5581 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5641 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5701 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5761 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5821 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5881 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5941 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
     6001 cccatatatg gagttccg
//
